中国兽医科学2024，Vol.54Issue(3)：350-356,7.DOI:10.16656/j.issn.1673-4696.2024.0047
非洲马瘟病毒NS1蛋白在昆虫细胞中的表达及抗原性分析
Expression of African horse sickness virus NS1 protein in insect cells and its antigenicity analysis
摘要
Abstract
African horse sickness(AHS)is a vector-borne viral disease of equids.To express the AHSV NS1 protein in baculovirus expression system and evaluate its antigenicity as a subunit vaccine component,based on the AHSV gene sequence published in GenBank,the full-length NS1 gene of AHSV was artificially synthesized.The complete open reading frame of NS1 gene was amplified by PCR and inserted into vector pFastBacTM HTb.The recombinant plasmid pFastBac-AHSV NS1 was transformed into DH10Bac to obtain recom-binant bacmid Bacmid-AHSV NS1.Bacmid-AHSV NS1 was transfected into Sf9 insect cells to rescue the recom-binant baculovirus expressing NS1 protein.SDS-PAGE and Western-blot showed that the molecular weight of recombinant AHSV NS1 protein was approximately 65 ku,and the purified recombinant AHSV NS1 protein was obtained.BALB/c mice was immunized with purified AHSV NS1 protein to obtain its polyclonal antibodies.Cellular immunofluorescence assay and Western-blot showed that the specific polyclonal antibodies against AHSV NS1 protein was obtained with a titer of 1∶52 000.Flow cytometry showed that the mice immunized with recombinant NS1 protein were able to induce the increase of CD4+and CD8+cells,indicating that AHSV NS1 protein has a potential application as a component of AHSV subunit vaccines.This study achieved the successful expression of AHSV NS1 protein in insect cells and verified its antigenicity,which laid a foundation for further investigation on the biological function of AHSV NS1 protein and the development of AHSV subunit vaccine.关键词
非洲马瘟病毒/AHSVNS1蛋白/昆虫细胞表达/抗原性Key words
African horse sickness virus/AHSV NS1 protein/insect cell expression/antigenicity分类
农业科技引用本文复制引用
户鑫兵,王轩莹,宋昱庆,田占成,关贵全,苟惠天,罗建勋,殷宏,独军政..非洲马瘟病毒NS1蛋白在昆虫细胞中的表达及抗原性分析[J].中国兽医科学,2024,54(3):350-356,7.基金项目
国家重点研发计划项目(2021YFD1800500) （2021YFD1800500）
中央级公益性科研院所基本科研业务费项目(1610312021014) （1610312021014）
甘肃省基础研究创新群体项目(22JR5RA024) （22JR5RA024）
甘肃省科技厅项目(22CX8NA011) （22CX8NA011）
农业科技创新工程项目(CAAS-ASTIP-2016-LVRI) （CAAS-ASTIP-2016-LVRI）
